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1 THE THREE PATHWAYS FOR THE REMOVAL OF AMMOIA-ITROGE I AQUACULTURE SYSTEMS James M. Ebeling, Ph.D. Research Engineer Aquaculture Systems Technologies, LLC Michael B. Timmons, Ph.D. Professor Dept. of Bio. & Environ. Eng. Cornell University

2 Introduction C inorganic / Ratio Ammonia-nitrogen C organic / Ratio Photoautotrophic (green-water systems) Autotrophic (fixed-cell bioreactors) Heterotrophic (zero-exchange systems) H 4+ - C 106 H 263 O P Algae H 4+ - O 2- - Ammonia Oxidizing Bacteria O 2- - O 3- - itrite Oxidizing Bacteria H 4+ - C 5 H 7 O 2 Bacteria

3 ew Paradigm??????? Understanding of the Removal System Photoautotrophic Autotrophic Heterotrophic Some Combination! Impact on Water Quality!!!! Management Strategies! Current Issues in Biofloc Technology Systems AAS-2011

4 itrogen Removal Pathways C organic H 4+ - CO 2 Alkalinity Trace utrients VSS Algae VSS Auto Extensive Pond O 2 Alkalinity CO 2 O 3- - Photoautotrophic Autotrophic Heterotrophic Other Mysterious Ways VSS Hetero

5 itrogen Removal Pathways C organic VSS Algae VSSAuto H 4+ - CO 2 Alkalinity Trace utrients Intensive Pond Alkalinity CO 2 Photoautotrophic Autotrophic Heterotrophic Other Mysterious Ways VSS Hetero O 2 O 3- - Algae Based Systems

6 itrogen Removal Pathways C organic H 4+ - O 2 Alkalinity Trace utrients Recirculation Systems Photoautotrophic Autotrophic Heterotrophic VSS Algae VSS Auto VSS Hetero CO 2 Alkalinity O 3- - O 2- - Denitrification Fixed-film Bioreactors

7 itrogen Removal Pathways C organic H 4+ - O 2 Alkalinity Trace utrients Zero-exchange Photoautotrophic Autotrophic Heterotrophic VSS Algae VSS Auto CO 2 Alkalinity O 3- - Denitrification VSS Hetero Suspended Growth Systems

8 Photoautotrophic (algal based systems) Biosynthesis of saltwater algae: itrate as nitrogen source 16 O CO H 2 O + HPO 2-4 C 106 H 263 O P O HCO - 3 Ammonia as nitrogen source 16 H CO H 2 O + 14 HCO 3- + HPO 4 2- C 106 H 263 O P O 2

9 Photoautotrophic (algal based systems) Consumes C organic C inorganic Consumables Stoichiometry (g) (g) (g) (g) H Carbon Dioxide g CO 2 / g Alkalinity 3.13 g Alk/ g Yields C organic C inorganic Products Stoichiometry (g) (g) (g) (g) VSS Algae g VSS A / g Oxygen g O 2 / g

10 Autotrophic - itrification Biosynthesis of Autotrophic bacteria: H O HCO C 5 H 7 O O H 2 O CO 2 The major factors affecting the rate of nitrification include: ammonia-nitrogen and nitrite-nitrogen concentration carbon/nitrogen ratio dissolved oxygen ph temperature alkalinity salinity

11 Autotrophic - itrification Consumes C organic C inorganic Consumables Stoichiometry (g) (g) (g) (g) H Alkalinity 7.05 g Alk/ g Oxygen 4.18 g O 2 / g Yields C organic C inorganic Products Stoichiometry (g) (g) (g) (g) VSS A 0.20 g VSS A / g O g O 3- - /g CO g CO 2 / g

12 Heterotrophic Bacteria Biosynthesis of Heterotrophic bacteria: H C 6 H 12 O 6 + HCO O 2 C 5 H 7 O H 2 O CO 2 The major factors affecting the rate of nitrification include: ammonia-nitrogen carbon/nitrogen ratio dissolved oxygen ph temperature alkalinity salinity

13 Heterotrophic Bacteria Consumes C organic C inorganic Consumables Stoichiometry (g) (g) (g) (g) H C 6 H 12 O g Carbs/ g Alkalinity 3.57 g Alk/ g Oxygen 4.71 g O 2 / g Yield C organic C inorganic Products Stoichiometry (g) (g) (g) (g) VSS H 8.07 g VSS H / g CO g CO 2 / g

14 Impact of C/ Ratio Autotrophic C labile / ~ 0 C organic / ~ small inorganic carbon as alkalinity 100 % Autotrophic (Recirculation System with excellent solids removal) C/ Ratio C labile / ~ 2.2 C/ ~ 8-10 organic carbon from the feed (109 g C organic /kg 35% protein 56 % Heterotrophic 44 % Autotrophic C labile / ~ 6.2 C/ ~ Heterotrophic organic carbon from the feed plus supplemental carbohydrates 100% Heterotrophic

15 Autotrophic (Intensive Recirculation System) Autotrophic System Conversion of 1 kg of 35% protein 32.2 g * 0.20 g VSS/ g = 6.44 g VSS autotrophic g/gvss A gc/gvss A 0.80 g VSS 3.42 g C VSS g O g CO 2 Current Issues in Biofloc Technology Systems AAS-2011

16 Autotrophic (Intensive Recirculation System) Consumes C organic C inorganic Consumables Stoichiometry (g) (g) (g) (g) H Alkalinity 7.05 g Alk/ g Oxygen 4.18 g O 2 / g Yields C organic C inorganic Products Stoichiometry (g) (g) (g) (g) VSS A 0.20 g VSS A / g O g O 3- - /g CO g CO 2 / g g C Alk / 32.2 g C/ ratio of 1.6 and TOC is very, very small Current Issues in Biofloc Technology Systems AAS-2011

17 Biofloc System (?) (no Carbon Supplementation) Heterotrophic and Autotrophic Components Heterotrophic Component: Organic Carbon from Feed 1 kg feed * 0.36 kg BOD/kg feed * 0.40 kg VSS/ kg BOD = = 144 g VSS heterotrophic g/gvss H gc/gvss H 17.9 g VSS 76.5 g C VSS g C CO2 = g C Assume that the heterotrophic bacteria out compete the autotrophic bacteria. 109 g C feed 15.4 g C Alkalinity Current Issues in Biofloc Technology Systems AAS-2011

18 Biofloc System (?) (no Carbon Supplementation) Heterotrophic and Autotrophic Components Autotrophic Component: Inorganic Carbon Alkalinity 14.3 g * 0.20 g VSS/ g Excess Ammonia-nitrogen: 32.2 g H g VSS = 14.3 g A = 2.86 g VSS autotrophic g/gvss A gc/gvss A 0.35 g VSS 1.52 g C VSS + 14 g O g C Alk

19 Biofloc System (?) (no Carbon Supplementation) Heterotrophic and Autotrophic Components Heterotrophic Bacteria Consumes C organic C inorganic Stoichiometry (g) (g) (g) (g) H * T C 6 H 12 O 6 feed g Carbs/ g Alkalinity 3.57 g Alk/ g Autotrophic Bacteria Consumes C organic C inorganic Stoichiometry (g) (g) (g) (g) H * T Alkalinity 7.05 g Alk/ g Conversion of 1 kg of 35% protein

20 Biofloc System (?) (no Carbon Supplementation) Heterotrophic and Autotrophic Components Heterotrophic Bacteria Yields C organic C inorganic Stoichiometry (g) (g) (g) (g) VSS H 8.07 g VSS H / g CO g CO 2 / g Autotrophic Bacteria Yields C organic C inorganic Stoichiometry (g) (g) (g) (g) VSS A 0.20 g VSS A / g O g O 3 -/g CO g CO 2 / g Conversion of 1 kg of 35% protein Current Issues in Biofloc Technology Systems AAS-2011

21 Biofloc System (?) (no Carbon Supplementation) Heterotrophic and Autotrophic Components C organic C inorganic Consumes (g) (g) (g) H g C 6 H 12 O g Carbs (feed) Alkalinity 165 g Alk C organic C inorganic Yields (g) (g) (g) VSS 147 g VSS O g O CO g CO Conversion of 1 kg of 35% protein Current Issues in Biofloc Technology Systems AAS-2011

22 Biofloc System (?) (no Carbon Supplementation) Heterotrophic and Autotrophic Components 100% 80% 97% 78% Heterotrophic Bacteria Autotrophic Bacteria Percent Removal 60% 40% 65% 56% 35% 22% 57% 51% 49% 44% 43% 39% 61% 35% 65% 20% 0% 3% 20% 25% 30% 35% 40% 45% 50% 55% Feed Protein (%) Percent removal of ammonia-nitrogen by Heterotrophic or Autotrophic Process as a function of % Protein Current Issues in Biofloc Technology Systems AAS-2011

23 Biofloc System Carbon Supplement Excess Ammonia-nitrogen: 32.2g H g VSS = 14.3g A 14.3 g * 8.07 g VSS / g = 115 g VSS heterotrophic g/gvss H gc/gvss H 14.3 g VSS 61.1 g C VSS g C CO2 = 98.7 g C Carbohydrate is 40% Carbon 216 g carbs 86.4 g C s 12.3 g C Alkalinity

24 Biofloc System (Carbon Supplementation) Consumes C organic C inorganic Consumables Stoichiometry (g) (g) (g) (g) H C 6 H 12 O g Carbs/ g Alkalinity 3.57 g Alk/ g Oxygen 4.71 g O 2 / g Yield C organic C inorganic Products Stoichiometry (g) (g) (g) (g) VSS H 8.07 g VSS H / g CO g CO 2 / g Conversion of 1 kg of 35% protein Current Issues in Biofloc Technology Systems AAS-2011

25 Biofloc System (Carbon Supplementation) Supplemental Carbohydrate. (% of feed). 60% 50% 40% 30% 20% 10% 0% 50% 43% 36% 29% 22% 15% 8% 0% 1% Feed Protein (%) Supplemental Carbohydrate as percentage of feed rate for heterotrophic metabolism of ammonia-nitrogen to microbial biomass Current Issues in Biofloc Technology Systems AAS-2011

26 Biofloc System (Carbon Supplementation) 500 VSS (g/kg feed) Feed Protein (%) VSS production as percentage of feed rate for heterotrophic metabolism of ammonia-nitrogen to microbial biomass Current Issues in Biofloc Technology Systems AAS-2011

27 Conclusions Engineering Sustainability Organic Carbon + itrogen Bacterial Biomass Current Issues in Biofloc Technology Systems AAS-2011

28 Questions? Current Issues in Biofloc Technology Systems AAS-2011

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